文献类型：期刊文献

中文题名：四海舒郁丸介导miRNA-494对甲状腺乳头状癌细胞的干预作用

英文题名：Study on the intervention effect of Sihaishuyu pill mediated by miRNA-494 on thyroid papillary carcinoma cells

作者：陈茜茜[1,2];伊琳[1,2];顾远晖[3];徐静[1,2];贾冠军[1,2];吕红英[1,2];候明双[1,2]

第一作者：陈茜茜

机构：[1]甘肃中医药大学,甘肃兰州730000;[2]甘肃省中医药防治慢性疾病重点实验室,甘肃兰州730000;[3]甘肃省人民医院,甘肃兰州730000

第一机构：甘肃中医药大学

年份：2024

卷号：59

期号：2

起止页码：8

中文期刊名：甘肃农业大学学报

外文期刊名：Journal of Gansu Agricultural University

收录：CSTPCD;;北大核心:【北大核心2023】；CSCD:【CSCD_E2023_2024】；

基金：甘肃省自然科学基金项目(20JR5RA177,20JR10RA368)。

语种：中文

中文关键词：四海舒郁丸;miRNA-494;甲状腺乳头状癌;PTEN;PI3K/AKT

外文关键词：Sihaishuyu pill;miRNA-494;papillary thyroid carcinoma;PTEN;PI3K/AKT

摘要：【目的】探讨四海舒郁丸含药血清对甲状腺乳头状癌细胞miRNA-494及靶基因PTEN和相关通路PI3K/AKT的干预机制。【方法】将2月龄SPF级雄性SD大鼠随机分为四海舒郁丸低、中、高剂量组与空白组,每组5只,灌胃给药后制备四海舒郁丸含药血清和空白组血清(等量生理盐水)并作用于体外培养的细胞。试验组甲状腺乳头状癌细胞选用KTC-1细胞株,分为空白组(2 mL/(100 g·d)生理盐水),低、中、高剂量组(分别为0.24、0.48、0.96 g/(100 g·d)),正常组甲状腺组织细胞选用Nthy-ori-3-1。通过CCK-8探究细胞的活力的影响;RTqPCR检测miRNA-494-5p及PTEN的mRNA表达水平,Western Blot法检测PTEN及下游PI3K/AKT通路蛋白表达水平。【结果】四海舒郁丸能抑制KTC-1细胞的活性,且作用随浓度的增加而增强(P<0.01),但无时间依赖性;相比较于KTC-1空白组,四海舒郁丸低、中、高剂量组中miRNA-494-5p的mRNA表达均降低(P<0.01),PTEN的mRNA表达量增加,其中高剂量组差异有统计学意义(P<0.05);高、中、低剂量组中p-PI3K/p-AKT信号通路蛋白表达均明显下降(P<0.05),而PTEN蛋白表达均有所增加(P<0.01)。【结论】四海舒郁丸含药血清通过下调miRNA-494-5p后负向调控PTEN进而阻断PI3K/AKT通路对PTC发挥辅助抑癌作用。
【Objective】To investigate the intervention mechanism of Sihaishuyu pill containing serum on miRNA-494,target gene PTEN and related signaling pathway PI3K/AKT in papillary thyroid carcinoma cells.【Method】SPF male Sprague-Dawley rats aged 2 months were randomly divided into low,medium,and high-dose Sihaishuyu pill group and blank control group,with 5 rats in each group.After gavage,the serum containing Sihaishuyu pill and the serum of the blank control group(with the same volume of normal saline)were prepared and used as cultured cells in vitro.Thyroid papillary cancer cells in the experimental group were divided into blank group(2 mL/(100g·d)normal saline),low,medium and high dose groups(0.24,0.48,0.96 g/(100g·d),respectively).Thyroid cells from the normal group were selected and Nthy-ori-3-1.CCK-8 was used to study the effect on cell viability.The mRNA expression levels of miRNA-494-5p and PTEN were detected by RT-qPCR,and the protein expression levels of PTEN and its downstream PI3K/AKT pathway were detected by Western blot.【Result】Sihaishuyu pill could inhibit the activity of KTC-1 cells in a dose-dependent manner(P<0.01),but not in a time-dependent manner.Compared with the KTC-1 blank group,the mRNA expression of miRNA-494-5p was decreased(P<0.01),and the mRNA expression of PTEN was increased in the low,medium and high dose groups of Sihaishuyu pills,and the difference in the high dose group was statistically significant(P<0.05).The expression of p-PI3K/p-Akt signaling pathway protein was significantly decreased in the high,medium and low dose groups(P<0.05),while the expression of PTEN protein was increased(P<0.01).【Conclusion】Sihaishuyu pill serum may have an additional anti-cancer effect on PTC by down-regulating miRNA-494-5P and negatively regulating PTEN to block the PI3K/AKT pathway.